Cephalosporins

ABSTRACT

A compound of the formula (I): ##STR1## wherein A is a monocyclic or polycyclic heteroaromatic ring containing at least one nitrogen atom as a hetero atom, which may be unsubstituted or substituted with one or more substituents; 
     R is a phenyl group which can be unsubstituted or substituted or a thienyl group; 
     T is (1) a --CH 2  --S--Het group, where Het is a tetrazolopyridazine ring, a triazolopyridazine ring, or a triazolopyridine ring, (2) a ##STR2##  group wherein R 1  and R 2 , which may be the same or different, each is a hydrogen atom or a (C 1  -C 4 )alkyl group, (3) a --CH 2  N 3  group or (4) is --CH 2  S--D group in which D is a group selected from the group consisting of ##STR3##  where m and n each is 0 to 3: with the proviso that (a) when the HO--A-- moiety is a ##STR4##  group where b represents the non-metallic atoms necessary to complete a pyridine ring, a pyrimidine ring or a pyrazole ring, each of which may be unsubstituted or substituted, then T is the --CH 2  SD group wherein D is as defined above, and 
     (b) when T is the --CH 2  N 3  group, then R is a substituted phenyl group 
      and the non-toxic, pharmaceutically acceptable salts thereof, processes for preparing the same, and anti-microbial compositions containing the same.

This is a division of application Ser. No. 773,729, filed Mar. 2, 1977now U.S. Pat. No. 4,165,373.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to novel cephalosporin compounds. Moreparticularly, this invention relates to cephalosporin compounds usefulas chemotherapeutic agents and particularly having Pseudomonas activityin addition to a broad antimicrobial spectral activity.

2. Description of the Prior Art

It is known that cephalosporin series compounds such as Cephalothin andCefazolin are very effective and are widely used as chemotherapeuticagents for infectious diseases caused by Gram-positive or Gram-negativebacteria.

However, these cephalosporin series compounds have no effect oninfectious diseases caused by Pseudomonas aeruginosa which have beenincreasingly spreading in recent years, and are often very difficult tocure. Cephalosporin series compounds which are effective againstPseudomonas aeruginosa are not yet commercially available.

SUMMARY OF THE INVENTION

As the result of various studies seeking a cephalosporin series compoundhaving a strong anti-Pseudomonas activity and a broad antimicrobialspectral activity, it has been found that cephalosporins of the formula(I) as described below and the pharmaceutically acceptable salts thereofhave a strong antimicrobial activity against Gram-positive as well asGram-negative bacteria including Pseudomonas aeruginosa and are usefulas antimicrobial agents for the treatment or the prevention ofinfectious diseases caused by Gram-negative or Gram-positive bacteria.

Particularly, the compounds of the invention exhibit a noticeableantimicrobial activity against bacteria to which known cephalosporinseries compounds are barely effective, such as Pseudomonas aeruginosa,indole positive Proteus, Serratia, Enterobacter aerogenus, andCephaloridine resistant E. coli.

Accordingly, an object of this invention is to provide novelcephalosporin compounds which are useful as antimicrobial agents.

In one embodiment, this invention provides cephalosporin compoundsrepresented by the formula (I): ##STR5## wherein A represents amonocyclic or polycyclic heteroaromatic ring containing at least onenitrogen atom as a hetero atom, which may be unsubstituted orsubstituted with one or more substituents; R represents a phenyl group,a substituted phenyl group or a thienyl group; T represents (1) a --CH₂--S--Het group wherein Het represents a tetrazolopyridazine group, atriazolopyridazine group or a triazolopyridine group, (2) a ##STR6##group wherein R₁ and R₂, which may be the same or different, eachrepresents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms,(3) a --CH₂ N₃ group or (4) a --CH₂ S--D-- group in which D represents agroup selected from the group consisting of ##STR7## wherein m and neach represents an integer of 0 to 3; with the proviso that

(a) when the HO-A- moiety is a ##STR8## group wherein B represents thenon-metallic atoms necessary to complete a pyridine ring, a pyrimidinering or a pyrazole ring, each of which may be unsubstituted orsubstituted, then T is the --CH₂ SD group wherein D is as defined above,and

(b) when T is the --CH₂ N₃ group, then R is a substituted phenyl group

and the non-toxic, pharmaceutically acceptable salts thereof.

In another embodiment of this invention, this invention providesprocesses for preparing the compounds of the formula (I) above.

In an even further embodiment of this invention, the invention providesan anti-microbial composition containing at least one compound of theformula (I) above.

DETAILED DESCRIPTION OF THE INVENTION

More specifically, the compounds of the present invention represented bythe formula (I) include the following compounds having the formulae(I-a), (I-b), (I-c) and (I-d): ##STR9## wherein A represents amonocyclic or polycyclic nitrogen-containing heteroaromatic group; Rrepresents a phenyl group, a substituted phenyl group or a thienylgroup; Het represents a tetrazolopyridazine group, a triazolopyridazinegroup or a triazolopyridine group and R₁ and R₂, which may be the sameor different, each represents a hydrogen atom or an alkyl group having 1to 4 carbon atoms; ##STR10## wherein R is as defined above; B representsthe non-metallic atoms necessary to complete a pyridine ring, apyrimidine ring or a pyrazole ring, each of which may be substituted orunsubstituted; and D represents ##STR11## wherein n and m are as definedabove; and ##STR12## wherein A is as defined above; R₃ represents ahydroxy group, a protected hydroxy group, an amino group, a ureido groupor a hydroxymethyl group; R₄ and R₅ each represents a member selectedfrom the group consisting of a hydrogen atom, a nitro group, adialkylamino group, an alkenoylamino group, an amino group, a hydroxygroup, an alkanoyloxy group, a lower alkyl group, a lower alkoxy group,a halogen atom, a trifluoromethyl group, a hydroxymethyl group and asulfamyl group.

In the above formulae (I), (I-a), (I-b) and (I-d), the heteroaromaticring represented by A may be, for example, naphthyridine,pyrazolopyridine, pyridopyrazine, pyridopyrimidine, pyridine,pyrimidine, pyridazine and triazine. These heteroaromatic rings for Amay be substituted with 1 to 4 substituents, of which examples are ahalogen (e.g., fluorine, chlorine, bromine and iodine) atom, a loweralkyl group, a lower alkoxy group, a lower alkanoyl group, a loweralkoxycarbonyl group, a lower alkylthio group, a mercapto group, ahydroxy group, a lower alkoxymethyl group, a cyano group, a nitro group,a lower alkylsulfonyl group, an arylsulfonyl group, a sulfamoyl group, acarbamoyl group, an aryloxycarbonylamino group, an acetoacetylaminogroup, a lower alkylamino group, a lower dialkylamino group, a lowerhaloalkyl group, a lower alkenyl group, an aryl group, a cycloalkylgroup, a cycloalkylene group, and a heterocyclic ring group containing 1or 2 nitrogen atoms.

With respect to the above groups and the moieties contained therein aswell as the groups and moieties to be described hereinafter, unlessotherwise indicated, the term "lower alkyl" preferably includes an alkylgroup or moiety having up to 4 carbon atoms; "lower alkoxy" preferablyincludes an alkoxy group or moiety having up to 4 carbon atoms; "loweralkanoyl" preferably includes an alkanoyl group or moiety having up to 5carbon atoms; "lower alkoxycarbonyl" preferably includes analkoxycarbonyl group or moiety having up to 5 carbon atoms; "loweralkylthio" preferably includes an alkylthio group or moiety having up to4 carbon atoms; "lower alkoxymethyl" preferably includes an alkoxymethylgroup or moiety having up to 5 carbon atoms; "lower alkylsulfonyl"preferably includes an alkylsulfonyl group or moiety having up to 4carbon atoms; "arylsulfonyl" preferably includes a phenylsulfonyl groupor moiety; "aryloxycarbonylamino" preferably includes aphenyloxycarbonylamino group or moiety; "lower alkylamino" preferablyincludes an alkylamino group or moiety having up to 4 carbon atoms;"lower dialkylamino" preferably includes a dialkylamino group or moietyof which each of the alkyl moieties thereof has up to 4 carbon atoms;"lower haloalkyl" preferably includes a chloro- or fluoro-substitutedalkyl group or moiety having up to 4 carbon atoms, for example,chloromethyl, trifluoromethyl, 2,2,2-trichloroethyl; "lower alkenyl"preferably includes an alkenyl group or moiety having up to 4 carbonatoms; "aryl" preferably includes a phenyl group or moiety; "cycloalkyl"preferably includes a cycloalkyl group or moiety having 3 to 6 carbonatoms; "cycloalkylene" preferably includes a cycloalkylene group ormoiety having 4 to 6 carbon atoms and "heterocyclic ring containing 1 or2 nitrogen atoms" preferably includes pyrrolidinyl, morpholyl,piperazinyl or piperidinyl.

In the formula (I), the hydroxy group on the heteroaromatic ring A ispreferably linked to a carbon atom adjacent the carbon atom to which the##STR13## moiety is connected.

In the above formulae, R represents a phenyl group which may beunsubstituted or substituted or a thienyl group and the term "a phenylgroup" which may be substituted as used with respect to R includes anoptionally substituted phenyl group of the formula: ##STR14## whereinR₆, R₇ and R₈, which may be the same or different, each represents ahydrogen atom, a nitro group, a lower dialkylamino (preferably, di-(C₁-C₄)alkylamino) group, a lower alkanoylamino (preferably, (C₂-C₅)alkanoylamino) group, a lower alkylsulfonamido (preferably, (C₁-C₄)alkylsulfonamido) group, an amino group, a hydroxy group, a loweralkanoyloxy (preferably, (C₂ -C₅)alkanoyloxy) group, a lower alkyl(preferably, (C₁ -C₄)alkyl) group, a lower alkoxy (preferably, (C₁-C₄)alkoxy) group, a chlorine atom, a bromine atom, a fluorine atom, aniodine atom, a trifluoromethyl group, a hydroxymethyl group, or asulfamyl group, preferably a hydrogen atom, a hydroxy group, a chlorineatom, a fluorine atom or a methoxy group.

The heterocyclic ring represented by the symbol Het in the --S--Hetgroup may be unsubstituted or substituted with one to four of a (C₁-C₄)alkyl group, a (C₁ -C₄)alkoxy group or a hydroxy group.

Examples of suitable groups for Het includetetrazolo[4,5-b]pyridazine-6-yl,3-hydroxypyridazino[3,2-c]-s-triazol-6-yl, pyrido[2,1-c]-s-triazol-3-yl,s-triazolo-[4,3-b]pyridazine-3-yl, s-triazolo[4,3-b]pyridazine-6-yl,3-methyl-s-triazolo[4,3-b]pyridazine-6-yl and the like.

Further, R₁ and R₂ each represents a hydrogen atom or a lower alkylgroup having 1 to 4 carbon atoms. Suitable examples of lower alkylgroups include a methyl group, an ethyl group, a propyl group, ann-butyl group, an isopropyl group, an isobutyl group and a t-butylgroup.

The fused heterocyclic ring ##STR15## can contain one or moresubstituents such as a lower (C₁ -C₄)alkyl group, a lower (C₁ -C₄)alkoxygroup, a lower (C₂ -C₅)alkanoyl group, a lower (C₂ -C₄)alkoxycarbonylgroup, a lower (C₁ -C₄)alkylthio group, a mercapto group, a hydroxygroup, a lower (C₁ -C₄)alkoxymethyl group, a halogen atom, a cyanogroup, a nitro group, a lower (C₁ -C₄)alkylsulfonyl group, anarylsulfonyl group, a sulfamoyl group, a carbamoyl group, anaryloxycarbonylamino group, an acetoacetylamine, a lower (C₁-C₄)alkylamino group, a lower di-(C₁ -C₄)alkylamino group, a halo-(C₁-C₄)alkyl group, an alkenyl group, an aryl group or a (C₃ -C₆)cycloalkylgroup.

Examples of suitble non-toxic pharmaceutically acceptable salts derivedfrom the compounds of formula (I) include the sodium salt, the potassiumsalt, the calcium salt, the magnesium salt, the triethylamine salt, thediethanolamine salt, the morpholine salt, the procaine salt, theL-arginine salt, and the L-lysine salt.

The α-carbon atom of the side chain (phenylglycine moiety) attached tothe 7-position of the formula (I) is an asymmetric carbon atom andtherefore two optically active isomers exist. These two isomers(D-diastereomer and L-diastereomer) and the DL-form are included withinthe scope of the present invention, but the D-diastereomer is preferred.

Preferred examples of the compounds of this invention are as follows.##STR16## wherein A is a divalent heteroaromatic ring selected from thegroup consisting of a naphthyridine ring, a pyrazolopyridine ring, apyridopyrazine ring, a pyridopyrimidine ring, a pyridine ring, and apyridazine ring, each of which can be unsubstituted or substituted witha (C₁ -C₄)alkylthio group; R is an unsubstituted group or a phenyl groupsubstituted with one or more substituents selected from the groupconsisting of a hydroxy group, an amino group and a thienyl group; Hetis a tetrazolopyridazine group, a triazolopyridazine group or atriazolopyridine group, each of which can be unsubstituted orsubstituted with a hydroxy group or a (C₁ -C₄)alkyl group; ##STR17##wherein A is a divalent heteroaromatic ring selected from the groupconsisting of a naphthyridine ring, a pyridopyrimidine ring and apyridine ring, each of which can be unsubstituted or substituted with a(C₁ -C₄)alkylthio group; R is a phenyl group which can be unsubstitutedor substituted with one or two substituents selected from the groupconsisting of a hydroxy group, an amino group and a chloro atom; R₁ andR₂, which can be the same or different, each is a hydrogen atom or a (C₁-C₄)alkyl group; ##STR18## wherein A is a naphthyridine ring, apyridopyrimidine ring or a pyrazolopyridine ring, each of which may beunsubstituted or substituted with a substituent selected from the groupconsisting of a (C₁ -C₄)alkyl group or a (C₁ -C₄)alkylthio group; R is aphenyl group which can be unsubstituted or substituted with one or moresubstituents selected from the group consisting of a hydroxy group, anamino group and a chlorine atom or a thienyl group; D is a groupselected from the group consisting of groups of the formula ##STR19##where m and n each is 0 to 3; ##STR20## wherein A is a naphthyridinering, a pyridine ring, a pyrimidine ring, a pyridazine ring or atriazine ring, each of which can be unsubstituted or substituted with a(C₁ -C₄)alkoxy group or a hydroxy group; R₃ is a hydroxy group, aprotected hydroxy group, an amino group, a ureido group or ahydroxymethyl group; R₄ and R₅, which can be the same or different, eachis a hydrogen atom, a nitro group, a di-(C₁ -C₄)alkylamino group, a (C₂-C₅)alkanoylamino group, an amino group, a hydroxy group, a (C₂-C₅)alkanoyloxy group, a (C₁ -C₄)alkyl group, a (C₁ -C₄)alkoxy group, ahalogen atom, a trifluoromethyl group, a hydroxymethyl group or asulfamyl group.

Among these cephalosporins, the following compounds are preferred.

(1)7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido)-α-(p-hydroxyphenyl)acetamido]-3-(tetrazolo[4,5-b]-pyridazine-6-ylthiomethyl)-3-cephem-4-carboxylicacid

and the non-toxic, pharmaceutically acceptable salts thereof

(2)7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido)-α-(p-hydroxyphenyl)acetamido]-3-(pyrido[2,1-c]-s-triazol-3-ylthiomethyl)-3-cephem-4-carboxylicacid

and the non-toxic, pharmaceutically acceptable salts thereof

(3)7-[D-α-(4-Hydroxypyridine-3-carboxamido)-α-(p-hydroxyphenyl)acetamido]-3-(tetrazolo[4,5-b]-pyridazine-6-ylthiomethyl)-3-cephem-4-carboxylicacid

and the non-toxic, pharmaceutically acceptable salts thereof

(4)7-[D-α-(4-Hydroxypyridine-3-carboxamido)-α-(p-hydroxyphenyl)acetamido]-3-carbamoyloxymethyl-3-cephem-4-carboxylicacid

and the non-toxic, pharmaceutically acceptable salts thereof

(5)7-[D-α-(4-Hydroxypyridine-3-carboxamido)-α-(m-aminophenyl)acetamido]-3-carbamoyloxymethyl-3-cephem-4-carboxylicacid

and the non-toxic, pharmaceutically acceptable salts thereof

(6)7-[D-α-(4-Hydroxypyridine-3-carboxamido)-α-phenylacetamido]-3-carbamoyloxymethyl-3-cephem-4-carboxylicacid

and the non-toxic, pharmaceutically acceptable salts thereof

(7)7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido)-α-(p-hydroxyphenyl)acetamido]-3-carbamoyloxymethyl-3-cephem-4-carboxylicacid

and the non-toxic, pharmaceutically acceptable salts thereof

(8)7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido)-α-phenylacetamido]-3-carbamoyloxymethyl-3-cephem-4-carboxylicacid

and the non-toxic, pharmaceutically acceptable salts thereof

(9)7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido-α-(m-aminophenyl)acetamido]-3-carbamoyloxymethyl-3-cephem-4-carboxylicacid

and the non-toxic, pharmaceutically acceptable salts thereof

(10) 7-[D-α-(2-Methylthio-5,8-dihydro-5-oxopyrido[2,3-d]-pyrimidine-6-carboxamido)-α-(p-hydroxyphenyl)acetamido]-3-carbamoyloxymethyl-3-cephem-4-carboxylicacid

and the non-toxic, pharmaceutically acceptable salts thereof

(11)7-[D-α-(5,8-Dihydro-5H-8-oxopyrido[3,2-d]pyrimidine-7-carboxamido)-.alpha.-(p-hydroxyphenyl)acetamido]-3-carbamoyloxymethyl-3-cephem-4-carboxylicacid

and the non-toxic, pharmaceutically acceptable salts thereof

(12)7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido-α-(3-chloro-4-hydroxyphenyl)acetamido]-3-carbamoyloxymethyl-3-cephem-4-carboxylicacid

and the non-toxic, pharmaceutically acceptable salts thereof

(13)7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido-α-(p-hydroxyphenyl)acetamido]-3-(5-carboxymethyl-1,3,4-thiadiazol-2-ylthiomethyl)-3-cephem-4-carboxylicacid

and the non-toxic, pharmaceutically acceptable salts thereof

(14)7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido-α-(p-hydroxyphenyl)acetamido]-3-(1-carboxymethyl-1,2,3,4-tetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid

and the non-toxic, pharmaceutically acceptable salts thereof

(15)7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido)-α-(p-hydroxyphenyl)acetamido]-3-azidomethyl-3-cephem-4-carboxylicacid

and the non-toxic, pharmaceutically acceptable salts thereof

The compounds of the formula (I) of the present invention can beprepared by the following methods:

The compounds of the formula (I) above can be prepared by reacting acarboxylic acid of the formula (II):

    HO--A--COOH                                                (II)

wherein A is as defined above, or a reactive derivative thereof, with acompound of the formula (III): ##STR21## wherein R and T are as definedabove, or a salt or derivative thereof.

Referring more particularly to this process, inert solvents which can beused in the reaction between the compounds of the formulae (II) and(III) include polar solvents such as dichloromethane, chloroform,acetone, tetrahydrofuran, dioxane, acetonitrile, methyl isobutyl ketone,ethyl alcohol, dimethylformamide, dimethylacetamide, dimethyl sulfoxide,nitromethane, hexamethylphosphoric triamide, sulfolane, and the like;non-polar solvents such as benzene, toluene, petroleum ether, n-hexaneand the like; and a mixture thereof. These solvents can also be used incombination with water.

The reactive derivatives of the compound (II) mean reactive derivativesof a carboxyl group, for example, an acid halide, an acid anhydride, anacid azolide, an active ester, an acid azide and the like. Referringmore particularly to these reactive derivatives, examples include mixedacid anhydrides or symmetric acid anhydrides with acids such as dialkylphosphoric acids, phenyl phosphoric acid, diphenyl phosphoric acid,dibenzyl phosphoric acid, halogenated phosphoric acids, dialkylphosphorous acids, sulfuric acid, methanesulfonic acid, toluenesulfonicacid, naphthalenesulfonic acid, alkylcarbonates, aliphatic carboxylicacids (for example, pivalic acid, pentanoic acid, isopentanoic acid,2-ethylbutanoic acid); acid azolides with imidazole, substitutedimidazoles, dimethylpyrazole, triazole, tetrazole, and the like; andactive esters such as cyanomethyl ester, methoxymethyl ester, vinylester, propargyl ester, p-nitrophenyl ester, 2,4-dinitrophenyl ester,trichlorophenyl ester, pentachlorophenyl ester, methanesulfonylphenylester, phenylazophenyl ester, phenylthiophenyl ester, p-nitrophenylthioester, p-cresolthio ester, carboxymethylthio ester, pyranyl ester,pyridyl ester, piperidyl ester, 8-quinolylthio ester, and esters withN,N'-dimethylhydroxylamine, 1-hydroxy-2-(1H)-pyridone,N-hydroxysuccinimide or N-hydroxyphthalimide.

Further, when the compounds of the formula (II) are used in the form ofthe free acid (or the salt thereof), it is preferred to carry out thereaction in the presence of coupling agents such asN,N'-dicyclohexylcarbodiimide,N-cyclohexyl-N-morpholinoethylcarbodiimide,N-cyclohexyl-N-(4-diethylaminocyclohexyl)carbodiimide,N,N'-diethylcarbodiimide, N,N'-diisopropylcarbodiimide,N-ethyl-N-(3-dimethylaminopropyl)carbodiimide,N,N'-carbonyldi(2-methylimidazole),pentamethyleneketene-N-cyclohexylimine,diphenylketene-N-cyclohexylimine, alkoxyacetylenes,1-alkoxy-1-chloroethylenes, trialkyl phosphites, polyphosphoric acidethyl ester, polyphosphoric acid isopropyl ester, phosphorusoxychloride, oxalylchloride, triphenylphosphine, diethylphosphonylazide, diphenylphosphonyl azide, 2-ethyl-7-hydroxybenzisoxazolium salts,2-ethyl-5-(m-sulfonyl)isoxazolium hydroxide inner salts,(chloromethylene)dimethyl ammonium chloride and the like.

Examples of salts of compounds of the formula (III) include an alkalimetal salt or an alkaline earth metal salt (for example, the sodium,potassium, calcium, etc., salts) of acids of the formula (III); organicamine salts (for example, trimethylamine, triethylamine, quinoline,collidine, etc., salts) of the acids of the formula (III); and organicsulfonic acid salts (for example, toluenesulfonic acid,naphthalenesulfonic acid, tetralinsulfonic acid, trifluoroacetic acid,hydrochloric acid, etc., salts) of the acids of the formula (III).

The derivatives of the compounds of the formula (III) can becarboxyl-protected derivatives in which the carboxyl group is protectedwith a conventional protecting group, and such derivatives may be in theform of the ester, amide or anhydride thereof.

Examples of these carboxyl-protected derivatives include a silyl ester,an organo-tin ester, a toluenesulfonyl ethyl ester, a p-nitrobenzylester, a benzyl ester, a phenacyl ester, a 2-furylmethyl ester, adiphenylmethyl ester, a substituted diphenylmethyl ester, ap-methoxybenzyl ester, a trityl ester, a benzoyloxymethyl ester, a loweralkanoyloxymethyl ester, a dimethylmethyleneamino ester, a p-nitrophenylester, a methylsulfonylphenyl ester, a methylthiophenyl ester, a t-butylester, a 4-picolyl ester, an iodoethyl ester, a trichloroethyl ester, aphthalimidomethyl ester, a 3,4-dimethoxy or 3,5-dimethylbenzyl ester, a2-nitrobenzyl ester, a 2,2'-dinitrobenzyl ester, an acetyloxycarbonylgroup, or a trichloroethyl ester thereof, and the compounds of theformula (III) in which the carboxyl group is protected with a group ofthe formula ##STR22## a group of the formula --N═CH--R' (in which R' isan alkyl group or an aryl group), or a group of the formula ##STR23##

In case of the silyl ester, other substituents of the compound of theformula (III), if any, such as a hydroxy group or an amino group may besilylated.

In case of these derivatives of compounds of the formula (III), theirhydrochloric acid, p-toluenesulfonic acid, naphthalene sulfonic acid ortetralin sulfonic acid salts may also be used.

The carboxyl-protecting group can be removed after the reaction undermild conditions, if necessary. For example, it can be removed by asolvolysis such as a hydrolysis and an alcoholysis, a catalytichydrogenation, a reduction, an oxidation, a nucleophilic substitutionreaction, a photochemical reaction or an enzymatic reaction.

The reaction between the acid of the formula (II) or the reactivederivative thereof and the compound of the formula (III) or thederivative thereof can generally be carried out at a temperature rangingfrom about -50° to about 50° C.

The starting material compounds of the formula (II) and their reactivederivatives are known compounds and can be prepared by known methods,e.g., as described in J. Am. Chem. Soc., 68, 1317 (1946); J. Chem. Soc.,(c), 1966, 1816; J. Chem. Soc., 1953, 4175; Helvetica Chimica Acta, 37,134 (1954); Chem. Pharm. Bull., 19 (7), 1482-6 (1971); J. Am. Chem.Soc., 78, 1938 (1956); J. Het. Chem., 9, 235 (1972); Roczuiki Chemii, 48(2) 321-4 (1974). The starting material compounds of the formula (III)wherein T is a group represented by --CH₂ --S--Het can be prepared in aconventional manner by reacting a compound of the formula (IV):##STR24## wherein Het is as defined above, with a compound of theformula (V): ##STR25## wherein R is as defined above.

This process can be carried out in substantially the same manner as theprocess of reacting the compound of the formula (II) with the compoundof the formula (III).

The compounds of the formula (III) wherein T is ##STR26## and R₁ and R₂are as defined above can be prepared by the method as described inGerman Patent Publication (DT-OS) No. 2,203,653.

The compounds of the formula (III) wherein T is --CH₂ --S--D (where D isas defined above) can be prepared by the method as described in JapanesePatent Application (OPI) Nos. 54580/76 and 88990/76.

The compounds of the formula (III) wherein T is --CH₂ N₃ can be preparedby the method as described in British Pat. No. 1,297,069.

An alternative method for preparing the compounds of the formula (I)comprises reacting an acylaminocarboxylic acid of the formula (VI):##STR27## wherein A and R are as defined above, or a reactive derivativethereof, with a compound of the formula (VII): ##STR28## wherein T is asdefined above, or a derivative thereof, to produce the compound of theformula (I).

The above reaction can be carried out under the reaction conditions setforth for the reaction between the compound of the formula (II) and thecompound of the formula (III).

The compounds of the formula (VI) described above are known compoundsand can be prepared by a conventional method (e.g., as described in U.S.Pat. No. 3,954,733).

The compounds of the formula (VII) as described above can be prepared byknown methods as described in Japanese Patent Application (OPI) No.58089/75, Japanese Patent Publication No. 5550/72, British Pat. No.1,297,069 and German Patent Publication (DT-OS) No. 2,203,653.

An alternative method for preparing the compounds of the formulae (I-a)and (I-c) comprises reacting an N-acylamino-α-arylacetamidocephalosporinof the formula (VIII): ##STR29## wherein A and R are as defined above,with a thiol compound of the formula (IX):

    HS--U                                                      (IX)

wherein U represents Het or D, each of which is as defined above, toproduce a compound of the formula (I-a) or (I-c): ##STR30## wherein A, Rand U are as defined above, using a conventional procedure as describedin, for example, Japanese Patent Publication Nos. 12136/71, 2340/71 and14734/71, Japanese Patent Application (OPI) No. 68593/73, J. Chem. Soc.,1965, p5015, etc.

For example, the reaction can be carried out in an inert solvent such aswater. Organic solvents such as acetone, acetonitrile, methanol,ethanol, dimethylformamide and the like may be used in combination withwater and a suitable buffer may also be used. When the compounds of theformula (VIII) are used in the form of the free carboxylic acid, thereaction preferably is carried out in the presence of a base such assodium bicarbonate or triethylamine. In general, the reaction ispreferably conducted at about 50° C. to about 60° C.

A still alternative method for preparing the compounds of the formulae(I-a) and (I-c) comprises reacting a compound of the formula (X):##STR31## wherein B and R are as defined above; Y represents a halogenatom or an ##STR32## moiety; and R" represents a hydrogen atom or anester group which is capable of being easily removed, with aheterocyclic thiol compound of the formula (IX):

    HS--U                                                      (IX)

wherein U is as defined above, as disclosed in, for example, JapanesePatent Application (OPI) No. 117487/74. That is, the reaction betweenthe compound of the formula (X) and the heterocyclic thiol compound canbe effected in a solvent such as dimethylformamide, hexamethylphosphorictriamide and the like, in the presence of a metal compound such ascupric chloride, cupric bromide, cupric fluoride, copper sulfate and thelike, at a temperature of about 0° C. to about 100° C.

Another method for preparing the compound of the formula (I-d) comprisesreacting an N-acylamino-α-arylacetamidocephalosporin of the formula(XI): ##STR33## wherein A, R₃, R₄ and R₅ are as defined above, withsodium azide (NaN₃) to produce the corresponding compound of the formula(I), as described, for example, in J. Chem. Soc., 1965, 5015. Morespecifically, the above reaction can be effected using a solvent such aswater, preferably a buffer solution, or a mixture thereof with anorganic solvent, e.g., as described above, at a temperature of about 50°C. to about 60° C.

Also, an alternative method for preparing the compound of the formula(I-b) comprises reacting a compound of the formula (XII): ##STR34##wherein A and R are as defined above, and E represents an ester residuewhich is capable of being easily removed, with a compound of the formula(XIII):

    XCONH.sub.2                                                (XIII)

wherein X represents a halogen atom, or a compound of the formula (XIV):

    ZSiNCO                                                     (XIV)

wherein Z represents a hydrogen atom or a methoxy group, to produce thecorresponding compound of the formula (I), as disclosed in, for example,Dutch Patent Publication Nos. 7,216,136 and 7,216,137.

The compounds of the formula (I) of this invention are valuable asantibacterial agents, nutritional supplements in animal feeds,therapeutic agents for poultry and animals, including man, and areespecially useful in the treatment of infectious diseases caused byGram-positive bacteria such as Staphylococcus aureus, Staphylococcusepidermidis, Staphylococcus pyogenes, Diplococcus pneumoniae, Sarcinalutea, Bacillus subtilis, Clostridium perfringens and Corynebacteriumdiphtheriae, and Gram-negative bacteria such as Escherichia coli,Neisseria gonouhoeae, Salmonella typhi, Klebsiella pneumoniae, Shigelladysenteriae, Shigella flexneri, Shigella sonnei, Enterobacter aerogenes,Proteus mirabilis, Proteus valgaris, Pseudomonas aeruginosa and Serratiamarcescens. For the treatment or prevention of such infectious diseases,the compounds of this invention, either individually or in combinationwith a pharmaceutically acceptable carrier or diluent, or another activeingredient(s), e.g., another chemotherapeutic agent(s), can beadministered intramuscularly or intravenously to a subject.

The dosage of the compounds of the formula (I) of this invention willvary with the body weight, age and conditions of an individual subject,the kind of bacteria, and the pharmacokinetic properties of theparticular compound chosen. Although the particular dosage will bedetermined by a physician taking these factors into consideration, thecompounds of the formula (I) are, in general, most desirablyadministered intramuscularly or intravenously at a dosage ranging fromabout 2 mg/kg of body weight/day to 400 mg/kg of body weight/day,preferably from 8 mg/kg of body weight/day to 120 mg/kg of bodyweight/day in a single dose or in multiple doses 1 to 5 times daily.

For intramuscular or intravenous administrations the compounds of thisinvention may be used in the form of sterile solution or suspensioncontaining additionally a pharmaceutically acceptable diluent or carriersuch as water, saline solution, Ringer's solution, glycerin,polyethylene glycol, etc. These preparations or formulations may alsocontain suitable auxiliary materials, such as stabilizers, buffersubstances, wetting agents, emulsifiers, local anesthetics, or saltsthat regulate the osmotic pressure. The compounds of the formula (I) ofthis invention may also be applied topically in the form of an ointmentor cream to the skin or other organs as a sterilizer or disinfectant.

The present invention is further illustrated in greater detail by thefollowing Examples and Reference Examples, but the invention is not tobe construed as being limited thereto. Unless otherwise indicated, allparts, percents and ratios are by weight.

EXAMPLE 1 Preparation of7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido)-α-p-hydroxyphenylacetamido]-3-(tetrazolo-[4,5-b]pyridazine-6-ylthiomethyl)-3-cephem-4-carboxylicAcid ##STR35##

1.23 g of sodium7-[D-α-(4-hydroxy-1,5-naphthyridine-3-carboxamido)-α-p-hydroxyphenylacetamido]-3-acetoxymethyl-3-cephem-4-carboxylate,0.20 g of sodium bicarbonate and 20 ml of a phosphate buffer (pH 6.4)were heated at a temperature of 60° C., and a solution of 0.416 g of6-mercaptotetrazolo[4,5-b]pyridazine in 10 ml of acetone was addeddropwise thereto. After completion of the dropwise addition, theresulting mixture was allowed to react for 12 hours and 20 minutes atthe same temperature. The thus obtained homogeneous reaction solutionwas then ice cooled whereby crystals precipitated. The crystals thusprecipitated were filtered, washed with 95% ethanol and dried overphosphorus pentoxide under reduced pressure to obtain 0.38 g of thetitled compound as the sodium salt.

Melting Point: 267°-274° C. (decomposition).

IR Absorption: ν_(Nujol) ^(cm).spsp.-1 1770, 1650, 1610.

EXAMPLE 2 Preparation of7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido)-α-p-hydroxyphenylacetamido]-3-(pyrido[2,1-c]-s-triazol-3-ylthiomethyl)-3-cephem-4-carboxylicAcid ##STR36##

1.23 g of sodium7-[D-α-(4-hydroxy-1,5-naphthyridine-3-carboxamido)-α-p-hydroxyphenylacetamido]-3-acetoxymethyl-3-cephem-4-carboxylate,0.45 g of sodium bicarbonate, 20 ml of a phosphate buffer (pH 6.4),0.906 g of 3-mercaptopyrido[2,1-c]-s-triazole and 20 ml of acetone werecharged into a reactor, and the resulting mixture was allowed to reactfor 23 hours at a temperature of 60° C. with stirring. The reactionsolution was then cooled and adjusted to a pH of 3.2 to 3.6 with 6 Nhydrochloric acid. After stirring the mixture under ice cooling, theprecipitated crystals were filtered, washed with water and dried overphosphorus pentoxide under reduced pressure to obtain 0.88 g of thetitled compound.

Melting Point: 221°-224° C. (decomposition).

IR Absorption: ν_(Nujol) ^(cm).spsp.-1 1760, 1655, 1610.

EXAMPLE 3 Preparation of7-[D-α-(4Hydroxypyridine-3-carboxamido)-α-p-hydroxyphenylacetamido]-3-(tetrazolo[4,5-b]pyridazine-6-ylthiomethyl)-3-cephem-4-carboxylicAcid ##STR37##

To a solution of 9 ml of dimethyl sulfoxide, 0.606 g of triethylamineand 0.472 g of 4-hydroxypyridine-3-carboxylic acid N-hydroxysuccinimideester was added 1.25 g of the trifluoroacetic acid salt of7-(D-α-amino-α-p-hydroxyphenylacetamido)-3-(tetrazolo[4,5-b]pyridazine-6-ylthiomethyl)-3-cephem-4-carboxylicacid, and the mixture was allowed to react for 12 minutes at roomtemperature (about 20°-30° C.). The reaction solution was then addeddropwise to 200 ml of acetone, and 100 ml of diethyl ether was furtheradded thereto whereby the crystals thus precipitated where filtered toobtain 0.99 g (yield: 70.2%) of the titled compound as the triethylaminesalt.

The resulting triethylamine salt was dissolved in 10 ml of water andthen the solution was adjusted to a pH of 3 with 2 N hydrochloric acidwith stirring under ice cooling. To the thus-obtained gel-like crystalswere further added 15 ml of water and 6 ml of methanol to thereby effectacid crystal formation. The precipitated crystals were filtered, washedwith methanol and dried over phosphorus pentoxide under reduced pressureto obtain 0.84 g of the titled compound.

The resulting compound was dissolved in 8 ml of dimethyl sulfoxide, and0.26 g of sodium 2-ethylhexanoate was added thereto followed by stirringthe mixture for 20 minutes at room temperature. The reaction solutionwas then added dropwise to 200 ml of acetone. The precipitated crystalswere filtered, washed successively with acetone and diethyl ether anddried over phosphorus pentoxide under reduced pressure to obtain 0.85 gof the titled compound as the sodium salt.

Melting Point: 253°-257° C. (decomposition).

IR Absorption: ν_(Nujol) ^(cm).spsp.-1 1760, 1660, 1630, 1610.

EXAMPLE 4 Preparation of7-[D-α-(4-Hydroxypyridine-3-carboxamido)-α-p-hydroxyphenylacetamido]-3-carbamoyloxymethyl-3-cephem-4-carboxylicAcid ##STR38##

4.22 g of7-(D-α-amino-α-p-hydroxyphenylacetamido)-3-(carbamoyloxymethyl)-3-cephem-4-carboxylicacid, 2.02 g of triethylamine and 30 ml of dimethyl sulfoxide werestirred at room temperature, and 2.36 g of4-hydroxypyridine-3-carboxylic acid N-hydroxysuccinimide ester was thenadded thereto whereby the mixture was allowed to react for 20 minutes atthe same temperature with stirring. Thereafter, 1.66 g of sodium2-ethylhexanoate was added to the resulting reaction mixture, and 10minutes later the reaction solution was added dropwise to 500 ml ofacetone. The precipitated titled compound as the sodium salt wasfiltered and dried over phosphorus pentoxide under reduced pressure toobtain 3.82 g of the dried product.

EXAMPLES 5 TO 12

The following compounds were synthesized in the same manner as describedin Example 4.

    ______________________________________                                         ##STR39##                                                                    Example No.                                                                              HOA              R                                                 ______________________________________                                                    ##STR40##                                                                                      ##STR41##                                        6                                                                                         ##STR42##                                                                                      ##STR43##                                        7                                                                                         ##STR44##                                                                                      ##STR45##                                        8                                                                                         ##STR46##                                                                                      ##STR47##                                        9                                                                                         ##STR48##                                                                                      ##STR49##                                        10                                                                                        ##STR50##                                                                                      ##STR51##                                        11                                                                                        ##STR52##                                                                                      ##STR53##                                        12                                                                                        ##STR54##                                                                                      ##STR55##                                        ______________________________________                                    

The results of the antimicrobial activities of these compounds in invitro testing in accordance with the recognized method are set forth inthe Table below.

                                      TABLE                                       __________________________________________________________________________    Minimum Inhibitory Concentration (μ g/ml)                                       Staphylococcus                                                                        Staphylococcus                                                                              Klebsiella                                                                          Proteus                                                                           Pseudomonas                              Example                                                                            aureus  aureus  Escherichia                                                                         pneumoniae                                                                          vulgaris                                                                          aeruginosa                               No.  209P    FS 289  coli NIHJ                                                                           602   HX 19                                                                             104                                      __________________________________________________________________________    4    0.39    3.13    12.5  12.5  0.2 6.25                                     5    0.39    3.13    6.25  12.5  0.2 6.25                                     6    0.39    3.13    12.5  12.5  0.2 12.5                                     7    0.78    6.25    3.13  1.56  0.78                                                                              6.25                                     8    0.78    6.25    3.13  1.56  0.78                                                                              6.25                                     9    0.78    3.13    1.56  1.56  0.78                                                                              6.25                                     10   0.78    3.13    6.25  6.25  1.56                                                                              6.25                                     11   1.56    6.25    6.25  6.25  1.56                                                                              12.5                                     12   1.56    6.25    3.13  1.56  0.78                                                                              6.25                                     __________________________________________________________________________

EXAMPLE 13 Preparation of7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido)-α-p-hydroxyphenylacetamido]-3-(5-carboxymethyl-1,3,4-thiadiazol-2-ylthiomethyl)-3-cephem-4-carboxylicAcid ##STR56##

1.23 g of sodium7-[D-α-(4-hydroxy-1,5-naphthyridine-3-carboxamido)-α-p-hydroxyphenylacetamido]-3-acetoxymethyl-3-cephem-4-carboxylate,0.20 g of sodium bicarbonate and 20 ml of a phosphate buffer (pH 6.4)were heated at a temperature of 60° C., and a solution of 0.48 g of(5-mercapto-1,3,4-thiadiazol-2-yl)acetic acid in 10 ml of acetone wasadded dropwise thereto. The resulting mixture was then allowed to reactfor 15 hours at the same temperature. The reaction solution was cooledand acidified with 6 N hydrochloric acid. The formed precipitate wasfiltered and washed with water to obtain the titled compound. Theresulting product was converted into the sodium salt using sodium2-ethylhexanoate in a usual manner.

IR Absorption: ν_(Nujol) ^(cm).spsp.-1 1770, 1650, 1610.

The antimicrobial activities of this compound were as follows.

    ______________________________________                                        Staphylococcus aureus 209 P                                                                       0.78 μg/ml                                             Escherichia coli NIHJ                                                                             1.56 μg/ml                                             Klebsiella pneumoniae 602                                                                         1.56 μg/ml                                             Pseudomonas aeruginosa 104                                                                        3.13 μg/ml                                             ______________________________________                                    

EXAMPLE 14 Preparation of7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido)-α-p-hydroxyphenylacetamido]-3-(1-carboxymethyl-1,2,3,4-tetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicAcid ##STR57##

The titled compound as the sodium salt was prepared in the same manneras described in Example 13 but using 5-mercapto-1H-tetrazol-1-aceticacid in place of (5-mercapto-1,3,4-thiadiazol-2-yl)acetic acid.

IR Absorption: ν_(Nujol) ^(cm).spsp.-1 1765, 1655, 1610.

The antimicrobial activities of this compound were as follows.

    ______________________________________                                        Staphylococcus aureus 209 P                                                                       0.78 μg/ml                                             Escherichia coli NIHJ                                                                             1.56 μg/ml                                             Klebsiella pneumoniae 602                                                                         1.56 μg/ml                                             Pseudomonas aeruginosa 104                                                                        3.13 μg/ml                                             ______________________________________                                    

EXAMPLE 15 Preparation of7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido)-α-p-hydroxyphenylacetamido]-3-azidomethyl-3-cephem-4-carboxylicAcid ##STR58##

A solution of 2.0 g of the sodium salt of7-[D-α-(4-hydroxy-1,5-naphthyridine-3-carboxamido)-α-p-hydroxyphenylacetamido]-3-acetoxymethyl-3-cephem-4-carboxylicacid, 0.328 g of sodium bicarbonate, 0.635 g of sodium azide and 40 mlof a phosphate buffer (pH 6.4) was allowed to react for 18 hours at atemperature of 55° C. with stirring. The insoluble matter was filteredout with heating, and the filtrate was then allowed to stand at roomtemperature whereby the turbidity formed was filtered out. The resultingreaction solution was then adjusted to a pH of 2 with 6 N hydrochloricacid, and the precipitated crystals were filtered and dried overphosphorus pentoxide under reduced pressure to obtain 0.80 g of thetitled compound.

This compound was dissolved in 5 ml of dimethyl sulfoxide, and theresulting mixture was reprecipitated with 30 ml of methanol to obtain0.55 g of the purified product.

The resulting product was further dissolved in 3 ml of dimethylsulfoxide, and 0.168 g of sodium 2-ethylhexanoate was added thereto.After stirring the mixture for 10 minutes, 30 ml of acetone was furtheradded dropwise thereto. The precipitated sodium salt was filtered,washed with acetone and dried over phosphorus pentoxide under reducedpressure to obtain 0.50 g of the titled compound as the sodium salt.

Melting Point: 283°-290° C. (decomposition).

IR Absorption: ν_(Nujol) ^(cm).spsp.-1 2100, 1765, 1650, 1610.

EXAMPLE 16 Preparation of7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido)-α-p-hydroxyphenylacetamido]-3-azidomethyl-3-cephem-4-carboxylicAcid

To a suspension of 1.43 g of 4-hydroxy-1,5-naphthyridine-3-carboxylicacid N-hydroxysuccinimide ester, 1.01 g of triethylamine and 30 ml ofdimethyl sulfoxide was added 2.11 g of7-(D-α-amino-p-hydroxyphenylacetamido)-3-aminocarbonyloxymethyl-3-cephem-4-carboxylicacid, and the mixture was allowed to react for 40 minutes at roomtemperature with stirring. A slight amount of insoluble materials wasthen filtered out, and the filtrate was added dropwise to 600 ml ofacetone. The precipitated crystals were filtered and dried overphosphorus pentoxide under reduced pressure to obtain 2.7 g of thetitled compound as the triethylamine salt. The sodium salt of the titledcompound was prepared using sodium 2-ethylhexanoate in the same manneras described in Example 15.

The resulting product was the same as that obtained in Example 15 andshowed the same IR absorption, NMR spectrum and Rf value (thin layerchromatography using a silica gel plate).

EXAMPLE 17 Preparation of7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido)-α-p-hydroxyphenylacetamido]-3-aminocarbonyloxymethyl-3-cephem-4-carboxylicAcid

1.70 g ofD-α-(4-hydroxy-1,5-naphthyridine-3-carboxamido)-α-p-hydroxyphenyl-aceticacid, which can be synthesized by the synthesis method as disclosed inU.S. Pat. No. 3,954,733, was dissolved in 60 ml of dimethyl sulfoxide,and 0.89 g of N,N'-carbonyldiimidazole was added to the resultingmixture at room temperature with stirring. 30 minutes later a solutionof 1.36 g of 7-amino-3-aminocarbonyloxymethyl-3-cephem-4-carboxylicacid, 0.76 g of triethylamine and 30 ml of dimethyl sulfoxide was addedthereto followed by allowing the mixture to react for 6 hours withstirring. The reaction solution was then added to 1.5 l of acetone, andthe precipitate formed was filtered. The filtered product was dissolvedin 50 ml of water and the solution adjusted to a pH of 2 with 3 Nhydrochloric acid under ice cooling. The thus formed precipitate wasfiltered, washed with water and dried over phosphorus pentoxide underreduced pressure to obtain 1.8 g of the titled compound. The sodium saltof this product which was obtained using sodium 2-ethylhexanoate wasconfirmed to contain the compound obtained in Example 7.

The following compounds can be also obtained in accordance with thepresent invention utilizing methods as described above.

    ______________________________________                                        Compounds of the Formula (I-a)                                                 ##STR59##                                                                    HOA              R          Het                                               ______________________________________                                         ##STR60##                                                                                      ##STR61##                                                                                ##STR62##                                         ##STR63##                                                                                      ##STR64##                                                                                ##STR65##                                         ##STR66##                                                                                      ##STR67##                                                                                ##STR68##                                         ##STR69##                                                                                      ##STR70##                                                                                ##STR71##                                         ##STR72##                                                                                      ##STR73##                                                                                ##STR74##                                         ##STR75##                                                                                      ##STR76##                                                                                ##STR77##                                         ##STR78##                                                                                      ##STR79##                                                                                ##STR80##                                         ##STR81##                                                                                      ##STR82##                                                                                ##STR83##                                         ##STR84##                                                                                      ##STR85##                                                                                ##STR86##                                         ##STR87##                                                                                      ##STR88##                                                                                ##STR89##                                         ##STR90##                                                                                      ##STR91##                                                                                ##STR92##                                         ##STR93##                                                                                      ##STR94##                                                                                ##STR95##                                         ##STR96##                                                                                      ##STR97##                                                                                ##STR98##                                         ##STR99##                                                                                      ##STR100##                                                                               ##STR101##                                       ______________________________________                                         *R configuration = Ddiastereomer                                         

    ______________________________________                                        Compounds of the Formula (I-b)                                                 ##STR102##                                                                    HOA                R                                                                                         ##STR103##                                    ______________________________________                                         ##STR104##                                                                                       ##STR105## NH.sub.2                                        ##STR106##                                                                                       ##STR107## NH.sub.2                                        ##STR108##                                                                                       ##STR109## NH.sub.2                                        ##STR110##                                                                                       ##STR111## NH.sub.2                                        ##STR112##                                                                                       ##STR113## NH.sub.2                                        ##STR114##                                                                                       ##STR115## NH.sub.2                                        ##STR116##                                                                                       ##STR117## NH.sub.2                                        ##STR118##                                                                                       ##STR119## NH.sub.2                                       ______________________________________                                         *R configuration = Ddiastereomer                                         

    __________________________________________________________________________    Compounds of the Formula (I-c)                                                 ##STR120##                                                                   HOA            R      Het                                                     __________________________________________________________________________     ##STR121##                                                                                   ##STR122##                                                                           ##STR123##                                              ##STR124##                                                                                   ##STR125##                                                                           ##STR126##                                              ##STR127##                                                                                   ##STR128##                                                                           ##STR129##                                              ##STR130##                                                                                   ##STR131##                                                                           ##STR132##                                              ##STR133##                                                                                   ##STR134##                                                                           ##STR135##                                              ##STR136##                                                                                   ##STR137##                                                                           ##STR138##                                              ##STR139##                                                                                   ##STR140##                                                                           ##STR141##                                              ##STR142##                                                                                   ##STR143##                                                                           ##STR144##                                              ##STR145##                                                                                   ##STR146##                                                                           ##STR147##                                              ##STR148##                                                                                   ##STR149##                                                                           ##STR150##                                              ##STR151##                                                                                   ##STR152##                                                                           ##STR153##                                             __________________________________________________________________________     *R configuration = Ddiastereomer                                         

    ______________________________________                                        Compounds of the Formula (I-d)                                                 ##STR154##                                                                    HOA                                                                                                  ##STR155##                                            ______________________________________                                         ##STR156##                                                                                           ##STR157##                                             ##STR158##                                                                                           ##STR159##                                             ##STR160##                                                                                           ##STR161##                                             ##STR162##                                                                                           ##STR163##                                             ##STR164##                                                                                           ##STR165##                                             ##STR166##                                                                                           ##STR167##                                             ##STR168##                                                                                           ##STR169##                                             ##STR170##                                                                                           ##STR171##                                             ##STR172##                                                                                           ##STR173##                                             ##STR174##                                                                                           ##STR175##                                             ##STR176##                                                                                           ##STR177##                                             ##STR178##                                                                                           ##STR179##                                            ______________________________________                                         *configuration = Ddiastereomer                                           

The following Reference Examples are given to illustrate the preparationof the starting materials used to prepare compounds of the presentinvention and the others are prepared in the same manner.

REFERENCE EXAMPLE 1 Preparation of7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido)-α-p-hydroxyphenylacetamido[-3-acetoxymethyl-3-cephem-4-carboxylicAcid

0.965 g of the trifluoroacetic acid salt of7-(D-α-amino-α-p-hydroxyphenylacetamido)-3-acetoxymethyl-3-cephem-4-carboxylicacid was dissolved in 8 ml of dimethyl sulfoxide, and 0.545 g oftriethylamine was added thereto. Subsequently, 0.516 g of4-hydroxy-1,5-naphthyridine-3-carboxylic acid N-hydroxysuccinimide esterwas added thereto followed by stirring the mixture for 1 hour and 15minutes at room temperature. 0.598 g of sodium 2-ethylhexanoate was thenadded to the resulting solution, and the mixture was stirred for 10minutes followed by filtering out the insoluble material. 100 ml ofacetone was added to the filtrate, and the precipitated crystals werethen filtered and dried over phosphorus pentoxide under reduced pressureto obtain 0.93 g of the titled compound as the sodium salt having amelting point (decomposition) of 261° to 265° C.

REFERENCE EXAMPLE 2 Preparation of7-[D-α-(4-Hydroxypyridine-3-carboxamido)-α-p-hydroxyphenylacetamido]-3-acetoxymethyl-3-cephem-4-carboxylicAcid

To a solution of 6 ml of dimethyl sulfoxide, 0.566 g of triethylamineand 0.441 g of 4-hydroxypyridine-3-carboxylic acid N-hydroxysuccinimideester was added 1 g of the trifluoroacetic acid salt of7-(D-α-amino-α-p-hydroxyphenylacetamido)-3-acetoxymethyl-3-cephem-4-carboxylicacid, and the mixture was allowed to react for 14 minutes at roomtemperature with stirring. The reaction solution was then added dropwiseto 250 ml of acetone, and 100 ml of diethyl ether was further addedthereto. The precipitated crystals were filtered and dried overphosphorus pentoxide under reduced pressure to obtain 0.99 g (yield82.5%) of the titled compound as the triethylamine salt having a meltingpoint of 135° C. for shrinking and 142° to 147° C. for decomposition.

This triethylamine salt was added to a solution of 0.31 g of sodium2-ethylhexanoate in 8 ml of dimethyl sulfoxide, and the mixture wasstirred for 10 minutes at room temperature. Thereafter, the resultingmixture was added to 180 ml of acetone and 50 ml of diethyl ether. Theprecipitated crystals were filtered and dried over phosphorus pentoxideunder reduced pressure to obtain the titled compound as the sodium salthaving a melting point (decomposition) of 150° to 165° C.

REFERENCE EXAMPLE 3 Preparation of7-Amino-3-(tetrazolo[4,5-b]pyridazine-6-ylthiomethyl)-3-cephem-4-carboxylicAcid ##STR180##

2.72 g of 7-aminocephalosporanic acid was suspended in 16 ml of waterand 8 ml of acetone, and a suspension of 2.1 g of sodium bicarbonate in16 ml of water was dropwise added thereto over a period of 7 minutestime. The inside temperature of a reactor was then increased to 50° C.,and a suspension of 2.3 g of 6-mercaptotetrazolo[4,5-b]pyridazine in 35ml of acetone was added dropwise thereto over a period of time of 10minutes. The resulting mixture was refluxed for 6 hours at a stableinside temperature of reactor of 61° C. followed by allowing therefluxed mixture to stand. The pH of the solution was 8 and thereafter,the mixture was adjusted to a pH of 3.6 with 6 N hydrochloric acid underice cooling, in which the pH was measured by means of a pH meter andthen stirred for 1 hour. The precipitated crystals were filtered anddried over phosphorus pentoxide under reduced pressure to obtain 2.39 gof the titled compound having a melting point (decomposition) of 202° to205° C.

REFERENCE EXAMPLE 4 Preparation of7-Amino-3-(s-triazolo[4,5-b]pyridazine-6-ylthiomethyl)-3-cephem-4-carboxylicAcid ##STR181##

2.72 g (0.01 mol) of 7-aminocephalosporanic acid was suspended in 20 mlof water and 10 ml of acetone, and a suspension of 1.89 g of sodiumbicarbonate in 10 ml of water was added dropwise thereto over a periodof time of 10 minutes. The inside temperature of the reactor was thenincreased to 50° C., and a suspension of 1.9 g (0.0125 mol) of2,3-triazolo-7,0-pyridazine-6-thiol in 20 ml of acetone was addeddropwise thereto over a period of time of 8 minutes. The resultingmixture was refluxed for 4 hours at a stable inside temperature of thereactor of 63° C. followed by ice cooling. After the inside temperatureof the reactor reached 3° to 4° C., the mixture was adjusted to a pH of5 with 6 N hydrochloric acid under ice cooling and then stirred for 10minutes to adjust the mixture to a pH of 3.6. One hour after thestirring, the precipitated crystals were filtered, washed with 10 ml ofwater and two times with 10 ml of acetone and dried over phosphoruspentoxide under reduced pressure to obtain 2.58 g (yield 72.5%) of thetitled compound having a melting point (decomposition) of 187° C.

Purification

A suspension of 1 g of the thus obtained crude product in 10 ml of waterwas stirred under ice cooling, and 9 ml of 12 N hydrochloric acid wasadded dropwise thereto incrementally over a period of time of 10minutes. Twenty minutes after the stirring, the insoluble material wasfiltered out, and the mother liquor was adjusted to a pH of 3.6 with 7ml of 30% sodium hydroxide and 3 ml of 10% sodium hydroxide. Thereafter,the resulting mixture was stirred for 1 hour, and the precipitatedcrystals were filtered and dried over phosphorus pentoxide under reducedpressure to obtain 0.84 g (yield: 84%) of the titled compound having amelting point (decomposition) of 194° C.

The same procedure as described above was followed using 10.74 g (0.0395mol) of 7-aminocephalosporanic acid, 134 ml of water, 7.5 g of sodiumbicarbonate and 7.5 g of 2,3-triazolo-7,0-pyridazine-6-thiol to obtain10.53 g (yield: 75%) of the titled compound as crude crystals. Thiscompound was found to have a melting point (decomposition) of 189° to192° C.

Subsequently, the same purification procedure described above wasfollowed using 10 g of the thus obtained crude product to obtain 8.2 g(yield: 82%) of the purified titled compound having a melting point(decomposition) of 194° C.

REFERENCE EXAMPLE 5 Preparation of7-Amino-3-(pyrido[2,1-c]-s-triazol-3-ylthiomethyl)-3-cephem-4-carboxylicAcid ##STR182##

2.72 g (0.01 mol) of 7-aminocephalosporanic acid was suspended in 18 mlof water, and a suspension of 2.1 g (0.025 mol) of sodium bicarbonate in20 ml of water was added dropwise thereto over a period of time of 10minutes. The inside temperature of the reactor was then increased to 50°C., and a suspension of 1.81 g (0.0125 mol) of3-mercaptopyrido[2,1-c]-s-triazole in 35 ml of acetone was addeddropwise thereto over a period of time of 10 minutes. The resultingmixture was refluxed for 4 hours at a stable inside temperature of thereactor of 63° C. followed by ice cooling the mixture. After the insidetemperature of the reactor reached 4° C., the mixture was adjusted to apH of 3.6 with 6 N hydrochloric acid. One hour after the stirring, theprecipitated crystals were filtered and dried over phosphorus pentoxideunder reduced pressure to obtain 2.31 g (yield: 61%) of the tiltedcompound having a melting point (decomposition) of 173° to 175° C.

REFERENCE EXAMPLE 6 Preparation of7-(D-α-amino-α-p-hydroxyphenylacetamido)-3-(tetrazolo[4,5-b]pyridazine-6-ylthiomethyl)-3-cephem-4-carboxylicAcid ##STR183##

3.31 g (0.01 mol) ofD(-)-N-p-methoxybenzyloxycarbonylamino-p-hydroxyphenylglycine wassuspended in 25 ml of dry acetonitrile, and 1.01 g of N-methylmorpholinewas added thereto. The resulting mixture was then cooled to -9° C.(inside temperature), and a solution of 1.36 g of chloroisobutylcarbonate in 2 to 3 ml of acetonitrile was added dropwise thereto. Theresulting mixture was stirred for 1.5 hours at a temperauture of -5° to-9° C. Separately, 4.01 g (0.011 mol) of7-amino-3-(tetrazolo-[4,5-b]pyridazine-6-ylthiomethyl)-3-cephem-4-carboxylicacid was suspended in 20 ml of dry acetonitrile, and 6.4 ml ofN,O-bis(trimethylsilyl) acetamide was added thereto followed by stirringthe mixture for 20 minutes at room temperature to obtain a disilylcompound containing solution. This disilyl compound containing solutionwas added at once to the above-described mixed anhydride solution, andthe mixture was allowed to react for 5 hours at a temperature of -10° to-2° C. After the mixture was further stirred for an additional 20minutes at a temperature of 0° to 8° C., 20 ml of methanol was addedthereto. Thus, the insoluble material was filtered out and the filtratewas then concentrated and dried to obtain an oil. To this oil were added1.68 g of sodium bicarbonate and 20 ml of water, and 20 ml of methanoland 40 to 50 ml of ethyl acetate were further added thereto to prepare ahomogeneous solution. The resulting solution was then concentrated underreduced pressure whereby the methanol was mainly discharged, and thesolution then separated into two liquid phases. The resulting aqueousphase was washed twice with 50 ml of ethyl acetate, and 50 ml of freshethyl acetate was then added thereover. The resulting solution wasadjusted to a pH of 2 to 1 with 6 N hydrochloric acid with stirringunder ice cooling. Thus, gummy materials were observed mixed therewith.The aqueous solution was extracted twice with ethyl acetate, and theresulting ethyl acetate layer was washed with a saturated sodiumchloride aqueous solution and dried over magnesium sulfate to obtain2.55 g of caramel-like crystals. Since these crystals contained a slightamount of D-N-p-methoxybenzyloxycarbonylamino-p-hydroxyphenylglycine,they were purified with ethyl acetate and diethyl ether to obtain 1.38 gof the desired compound. Further, when the gummy materials were mixedwith normal butanol, crystallization took place. The resulting compoundwas confirmed to be 2.0 g of the fairly pure desired compound. This,with the 1.38 g obtained as described above, resulted in 3.98 g of thedesired compound being obtained in a yield of 58.7%. ##STR184##

To an ice cooled solution of 16 ml of trifluoroacetic acid and 4 ml ofanisole was added 2.4 g of the above-described compound, and the mixturewas stirred for 35 minutes at room temperature. Thereafter, theresulting solution was added dropwise to 350 ml of diethyl ether. Theprecipitate formed was filtered, washed with diethyl ether and driedover phosphorus pentoxide under reduced pressure to obtain 1.94 g of thetitled compound in a yield of 87.5%.

While the invention has been described in detail and with reference tospecific embodiments thereof, it will be apparent to one skilled in theart that various changes and modifications can be made therein withoutdeparting from the spirit and scope thereof.

What is claimed is:
 1. A compound of the formula (I-b): ##STR185## wherein A is a divalent heteroaromatic ring selected from the group consisting of a naphthyridine ring, a pyridopyrimidine ring and a pyridine ring, each of which can be unsubstituted or substituted with a (C₁ -C₄)alkyl group or a (C₁ -C₄)alkylthio group; R is a phenyl group which can be unsubstituted or substituted with one or two substituents selected from the group consisting of a hydroxy group, an amino group and a chlorine atom; R₁ and R₂, which can be the same or different, each is a hydrogen atom or a (C₁ -C₄)alkyl group.
 2. A compound of the formula (I-b): ##STR186## wherein A is a divalent heteroaromatic ring selected from the group consisting of a naphthyridine ring, a pyridopyrimidine ring and a pyridine ring, each of which can be unsubstituted or substituted with a (C₁ -C₄)alkylthio group; R is a phenyl group which can be unsubstituted or substituted with one or two substituents selected from the group consisting of a hydroxy group, an amino group and a chlorine atom; and R₁ and R₂ each is a hydrogen atom.
 3. 7-[D-α-(4-Hydroxypyridine-3-carboxamido)-α-(p-hydroxyphenyl)acetamido]-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid and the non-toxic, pharmaceutically acceptable salts thereof, of claim
 1. 4. 7-[D-.alpha.(4-Hydroxypyridine-3-carboxamido)-α-(m-aminophenyl)acetamido]-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid and the non-toxic, pharmaceutically acceptable salts thereof, of claim
 1. 5. 7-[D-α-(4-Hydroxypyridine-3-carboxamido)-.alpha.-phenylacetamido]-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid and the non-toxic, pharmaceutically acceptable salts thereof, of claim
 1. 6. 7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido)-.alpha.-(p-hydroxyphenyl)acetamido]-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid and the non-toxic pharmaceutically acceptable salts thereof, of claim
 1. 7.7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido)-α-phenylacetamido]-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid and the non-toxic, pharmaceutically acceptable salts thereof, of claim
 1. 8. 7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido)-α-(m-aminophenyl)acetamido]-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid and the non-toxic, pharmaceutically acceptable salts thereof, of claim
 1. 9. 7-[D-.alpha.-(2-Methylthio-5,8-dihydro-5-oxopyrido[2,3-d]pyrimidine-6-carboxamido)-.alpha.-(p-hydroxyphenyl)-acetamido]-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid and the non-toxic, pharmaceutically acceptable salts thereof, of claim
 1. 10. 7-[D-α-(5,8-Dihydro-5H-8-oxopyrido[3,2-d]-pyrimidine-7-carboxamido)-.alpha.-(p-hydroxyphenyl)acetamdio]-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid and the non-toxic, pharmaceutically acceptable salts thereof, of claim
 1. 11. 7-[D-α-(4-Hydroxy-1,5-naphthyridine-3-carboxamido)-α-(3-chloro-4-hydroxyphenyl)acetamido]-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid and the non-toxic, pharmaceutically acceptable salts thereof, of claim
 1. 12. The sodium salt of the acid of claim
 6. 13. The potassium salt of the acid of claim
 6. 14. The acid of claim
 6. 15. A non-toxic, pharmaceutically acceptable salt of the acid of claim
 6. 16. The sodium salt of the acid of claim
 7. 17. The potassium salt of the acid of claim
 7. 18. The acid of claim
 7. 19. A non-toxic, pharmaceutically acceptable salt of the acid of claim
 7. 20. An acid having the D configuration in the 7-side-chain and the formula ##STR187## wherein R is hydrogen or hydroxy.
 21. An acid having the D configuration in the 7-side-chain and the formula ##STR188## wherein R is hydrogen or hydroxy or a non-toxic, pharmaceutically acceptable salt of said acid.
 22. The sodium salt of the acid of claim
 21. 23. The potassium salt of the acid of claim
 21. 24. A non-toxic, pharmaceutically acceptable salt of the acid of claim
 21. 25. An antibacterial composition which comprises an antibacterially effective amount of a compound of claim 1 as an active ingredient and a pharmaceutically acceptable carrier or diluent. 